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PMID:11563969

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Citation

Suginta, W, Karoulias, N, Aitken, A and Ashley, RH (2001) Chloride intracellular channel protein CLIC4 (p64H1) binds directly to brain dynamin I in a complex containing actin, tubulin and 14-3-3 isoforms. Biochem. J. 359:55-64

Abstract

Mammalian chloride intracellular channel (CLIC) (p64-related) proteins are widely expressed, with an unusual dual localization as both soluble and integral membrane proteins. The molecular basis for their cellular localization and ion channel activity remains unclear. To help in addressing these problems, we identified novel rat brain CLIC4 (p64H1) binding partners by affinity chromatography, mass spectrometric analysis and microsequencing. Brain CLIC4 binds dynamin I, alpha-tubulin, beta-actin, creatine kinase and two 14-3-3 isoforms; the interactions are confirmed in vivo by immunoprecipitation. Gel overlay and reverse pull-down assays indicate that the binding of CLIC4 to dynamin I and 14-3-3zeta is direct. In HEK-293 cells, biochemical and immunofluorescence analyses show partial co-localization of recombinant CLIC4 with caveolin and with functional caveolae, which is consistent with a dynamin-associated role for CLIC4 in caveolar endocytosis. We speculate that brain CLIC4 might be involved in the dynamics of neuronal plasma membrane microdomains (micropatches) containing caveolin-like proteins and might also have other cellular roles related to membrane trafficking. Our results provide the basis for new hypotheses concerning novel ways in which CLIC proteins might be associated with cell membrane remodelling, the control of cell shape, and anion channel activity.

Links

PubMed PMC1222121

Keywords

14-3-3 Proteins; Actins/metabolism; Amino Acid Sequence; Animals; Brain/metabolism; Caveolae/metabolism; Caveolin 1; Caveolins/metabolism; Cell Membrane; Cells, Cultured/cytology; Cells, Cultured/metabolism; Chloride Channels/metabolism; Chromatography, Affinity; DNA Primers/chemistry; Dynamin I/metabolism; Endocytosis; Glutathione Transferase/metabolism; Humans; Ion Channels/physiology; Mass Spectrometry; Molecular Sequence Data; Neurons/metabolism; Peptide Fragments/chemistry; Polymerase Chain Reaction; Precipitin Tests; Rats; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Tubulin/metabolism; Tyrosine 3-Monooxygenase/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:1433Z

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:61857

F

Seeded From UniProt

complete

RAT:1433Z

enables

GO:0044877: protein-containing complex binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:61857
RGD:71096

F

Seeded From UniProt

complete

RAT:1433Z

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:CLIC4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:3980

F

Seeded From UniProt

complete

RAT:CLIC4

enables

GO:0044877: protein-containing complex binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:3980
RGD:71096

F

Seeded From UniProt

complete

RAT:DYN1

enables

GO:0044877: protein-containing complex binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:3980
RGD:61857

F

Seeded From UniProt

complete

See also

References

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